Fig. 5.

Neo1^Gt/Gt MEFs are sensitized to SHH- and SAG-mediated Gli1 induction but neogenin does not bind SHH. A: qRT- PCR analysis of Gli1 and Ptch1 expression in Neo1^+/+ and Neo1^Gt/Gt MEFs. Two independent isolates of Neo1^+/+ MEFs and three independent isolates of Neo1^Gt/Gt MEFs were analyzed in duplicate in three separate experiments. Data were normalized to expression of Gapdh presented as fold induction over control, PBS-treated WT MEFs. Data represent means ± S.E.M. and were analyzed by Student’s t-test. *, p < 0.02; **, p < 0.005. B: Recombinant, secreted proteins comprising the ectodomains of neogenin or the SHH co-receptor, CDO, fused in-frame with the Fc region of human IgG (Neo1-Fc and CDO-Fc, respectively) were bound to protein-A sepharose. SHH-N::AP or, as a control, AP itself, were allowed to bind the Neo1-Fc and CDO-Fc matrices, which were then washed exhaustively. Bound AP activity was quantified with AP yellow liquid substrate.